■知窍甘草汤(ZQGCD)由龚正峰教授创作,著名的中医专家。临床研究表明,其在减轻椎间盘退变(IDD)患者的疼痛和增强腰椎功能方面具有功效。然而,ZQGCD治疗IDD的确切机制尚不清楚.
■使用液相色谱-串联质谱法(LC-MS/MS)鉴定ZQGCD的活性组分。建立大鼠椎间盘退变模型,每组大鼠接受ZQGCD治疗3周。评估参数包括痛觉过敏状态,观察椎间盘组织退变和巨噬细胞浸润,并剖析JAK2/STAT3通路卵白在椎间盘中的表达情形。用LPS诱导初级巨噬细胞M1极化,用JAK2抑制剂(AZD1480)和ZQGCD处理的细胞评估巨噬细胞极化,细胞上清液炎症因子,和JAK2/STAT3通路表达。以巨噬细胞上清液为条件培养基,观察其对髓核细胞(NPCs)增殖及胶原Ⅱ和MMP3蛋白表达的影响。
■在ZQGCD中鉴定出总共81种活性组分。ZQGCD治疗后,模型大鼠椎间盘组织中浸润的巨噬细胞减少,M1巨噬细胞的含量下降,而M2巨噬细胞的含量增加,血清促炎因子和疼痛诱导因子的表达降低,椎间盘组织中P物质的表达减少。因此,改善了大鼠椎间盘退变和痛觉过敏。体外研究表明,LPS诱导M1巨噬细胞极化。通过抑制JAK2/STAT3通路,JAK2抑制剂和ZQGCD均有效抑制M1极化,导致IL-1β水平降低,IL-6,TNF-α,和各种其他炎症因子。因此,这种抑制作用导致NPCs变性的延迟。
■IDD大鼠椎间盘组织有巨噬细胞浸润,JAK2/STAT3通路被激活,巨噬细胞极化为M1型,导致炎症微环境,导致椎间盘退变和痛觉过敏。ZQGCD通过抑制JAK2/STAT3/巨噬细胞M1极化途径表现出对IDD的延缓作用和改善痛觉过敏。
UNASSIGNED: Zhiqiao Gancao decoction (ZQGCD) was created by Professor Gong Zhengfeng, a renowned Chinese medicine expert. Clinical studies have shown its efficacy in alleviating pain and enhancing lumbar function in intervertebral disc degeneration (IDD) patients. However, the precise mechanism of ZQGCD in treating IDD remains unclear.
UNASSIGNED: The active components of ZQGCD were identified using Liquid chromatography-tandem mass spectrometry (LC-MS/MS). A rat model of intervertebral disc degeneration was established, and rats in each group received ZQGCD for three weeks. Assessment parameters included hyperalgesia status, observation of intervertebral disc tissue degeneration and macrophage infiltration, and analysis of JAK2/STAT3 pathway protein expression in the intervertebral disc. Primary macrophage M1
polarization was induced using LPS, with cells treated using the JAK2 inhibitor (AZD1480) and ZQGCD to evaluate macrophage
polarization, cellular supernatant inflammatory factors, and JAK2/STAT3 pathway expression. Macrophage supernatant served as a conditioned medium to observe its effects on the proliferation of nucleus pulposus cells (NPCs) and the expression of collagen II and MMP3 proteins.
UNASSIGNED: A total of 81 active components were identified in ZQGCD. Following ZQGCD treatment, infiltrating macrophages in intervertebral disc tissues of model rats decreased, the content of M1 macrophages decreased, while the content of M2 macrophages increased, the expression of proinflammatory factors and pain-inducing factors in serum decreased, and the expression of substance P in intervertebral disc tissue decreased. Consequently, the intervertebral disc degeneration and hyperalgesia of rats were improved. In vitro studies revealed that LPS induced M1 macrophage
polarization. By inhibiting the JAK2/STAT3 pathway, both JAK2 inhibitors and ZQGCD effectively suppressed M1
polarization, resulting in decreased levels of IL-1β, IL-6, TNF-α, and various other inflammatory factors. Consequently, this inhibition led to a delay in the degeneration of NPCs.
UNASSIGNED: There is macrophage infiltration in the intervertebral disc tissue of IDD rats, and JAK2/STAT3 pathway is activated, macrophages are polarized to M1 type, resulting in inflammatory microenvironment, leading to intervertebral disc degeneration and hyperalgesia. ZQGCD exhibited a delaying effect on IDD and improved hyperalgesia by inhibiting the JAK2/STAT3/macrophage M1
polarization pathway.